DE502004004202D1 - METHOD FOR DETECTING CYTOSINE METHYLATION IN DNA - Google Patents

METHOD FOR DETECTING CYTOSINE METHYLATION IN DNA

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Publication number
DE502004004202D1
DE502004004202D1 DE502004004202T DE502004004202T DE502004004202D1 DE 502004004202 D1 DE502004004202 D1 DE 502004004202D1 DE 502004004202 T DE502004004202 T DE 502004004202T DE 502004004202 T DE502004004202 T DE 502004004202T DE 502004004202 D1 DE502004004202 D1 DE 502004004202D1
Authority
DE
Germany
Prior art keywords
dna
base
probe
hybridization
detecting
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
DE502004004202T
Other languages
German (de)
Inventor
Reimo Tetzner
Juergen Distler
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Epigenomics AG
Original Assignee
Epigenomics AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Epigenomics AG filed Critical Epigenomics AG
Publication of DE502004004202D1 publication Critical patent/DE502004004202D1/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/154—Methylation markers

Abstract

Detecting cytosine methylation in DNA (I) by reacting (I) with an enzyme or chemical so that 5-methylcytosine (5MC) remains unchanged, while non-methylated cytosine (C) is converted to uracil or some other base having base-pairing properties different from those of C, is new. Detecting cytosine methylation in DNA (I) comprises: (a) reacting (I) with an enzyme or chemical so that 5-methylcytosine (5MC) remains unchanged, while non-methylated cytosine (C) is converted to uracil or some other base having base-pairing properties different from those of C; (b) amplifying the pretreated (I) using a polymerase and at least one primer that is linked at its 5'-end to a probe (scorpion primer, SP); (c) separating the primer extension product (PEP) from the matrix strand; (d) intracellular hybridization of the probe to PEP, where hybridization is dependent on the methylation status of the DNA; and (e) detecting whether or not hybridization of the probe has occurred. An independent claim is also included for a kit comprising SP, polymerase and reagents for performing a PCR.
DE502004004202T 2003-08-15 2004-08-13 METHOD FOR DETECTING CYTOSINE METHYLATION IN DNA Active DE502004004202D1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10338308A DE10338308B4 (en) 2003-08-15 2003-08-15 Method for the detection of cytosine methylations in DNA
PCT/DE2004/001837 WO2005024056A2 (en) 2003-08-15 2004-08-13 Method for the detection of cytosine methylations in dna with the aid of scorpion primers

Publications (1)

Publication Number Publication Date
DE502004004202D1 true DE502004004202D1 (en) 2007-08-09

Family

ID=34201746

Family Applications (2)

Application Number Title Priority Date Filing Date
DE10338308A Expired - Fee Related DE10338308B4 (en) 2003-08-15 2003-08-15 Method for the detection of cytosine methylations in DNA
DE502004004202T Active DE502004004202D1 (en) 2003-08-15 2004-08-13 METHOD FOR DETECTING CYTOSINE METHYLATION IN DNA

Family Applications Before (1)

Application Number Title Priority Date Filing Date
DE10338308A Expired - Fee Related DE10338308B4 (en) 2003-08-15 2003-08-15 Method for the detection of cytosine methylations in DNA

Country Status (9)

Country Link
US (1) US9863001B2 (en)
EP (2) EP1654388B1 (en)
JP (1) JP4743787B2 (en)
AT (1) ATE365813T1 (en)
CA (1) CA2540378A1 (en)
DE (2) DE10338308B4 (en)
DK (1) DK1654388T3 (en)
ES (1) ES2289594T3 (en)
WO (1) WO2005024056A2 (en)

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JP5133238B2 (en) 2005-04-15 2013-01-30 エピゲノミックス アクチェンゲゼルシャフト Method for providing a DNA fragment from a remote sample
BRPI0712655A2 (en) * 2006-06-08 2012-11-20 Koninkl Philips Electronics Nv microelectronic sensor device, method for investigating at least one biologically target substance, and, use of microelectronic sensor device
JP2008136404A (en) * 2006-11-30 2008-06-19 Sysmex Corp Method for confirming amount of dna after conversion treatment of non-methylated cytosine in dna methylation detection
US20090203011A1 (en) * 2007-01-19 2009-08-13 Epigenomics Ag Methods and nucleic acids for analyses of cell proliferative disorders
EP2152902A2 (en) 2007-06-08 2010-02-17 Epigenomics AG Method for methylation analysis
NZ583796A (en) * 2007-09-17 2011-12-22 Oncomethylome Sciences Sa Improved detection of mage-a expression
AU2008334901A1 (en) 2007-12-11 2009-06-18 Epigenomics Ag Methods and nucleic acids for analyses of lung carcinoma
EP2071035A1 (en) * 2007-12-13 2009-06-17 Epigenomics AG Facilitator and method for amplification
FR2940805B1 (en) 2009-01-05 2015-10-16 Biomerieux Sa METHOD FOR AMPLIFYING AND / OR DETECTING NUCLEIC ACIDS, KITS AND USES THEREOF
PL2210954T3 (en) * 2009-01-22 2011-12-30 Heinrich Heine Univ Duesseldorf Determining of the DNA methylation level
US20110117546A1 (en) * 2009-11-18 2011-05-19 Microfluidic Systems, Inc. Increase of signal sensitivity using dual probes in pcr reactions
WO2013084075A2 (en) * 2011-12-06 2013-06-13 Mdxhealth Sa Methods of detecting mutations and epigenetic changes
EP3033445B1 (en) * 2013-08-12 2020-01-01 Bio-Rad Laboratories, Inc. Amplification reporter with base-pairing oligomers
GB201402644D0 (en) * 2014-02-14 2014-04-02 Base4 Innovation Ltd Methylation detection method
CN109804086B (en) 2016-08-10 2023-06-13 格里尔公司 Method for preparing double-tag DNA library for sequencing sulfite transformation
CN108456721B (en) * 2018-05-16 2021-07-09 基因科技（上海）股份有限公司 Method for synchronously detecting gene mutation and methylation and application thereof
US11427874B1 (en) 2019-08-26 2022-08-30 Epi One Inc. Methods and systems for detection of prostate cancer by DNA methylation analysis
WO2021192395A1 (en) 2020-03-25 2021-09-30 富士フイルム株式会社 Method and program for calculating base methylation levels
CN113552191B (en) * 2021-07-28 2023-11-21 江苏师范大学 Construction method of proportional electrochemical sensor for detecting methylated DNA based on multilayer DNA amplification loop
CN114317761A (en) * 2022-02-23 2022-04-12 厦门飞朔生物技术有限公司 Methylation detection method for cervical cancer related gene

Family Cites Families (20)

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Publication number Priority date Publication date Assignee Title
GB8920097D0 (en) 1989-09-06 1989-10-18 Ici Plc Amplification processes
US5786146A (en) 1996-06-03 1998-07-28 The Johns Hopkins University School Of Medicine Method of detection of methylated nucleic acid using agents which modify unmethylated cytosine and distinguishing modified methylated and non-methylated nucleic acids
US6017704A (en) 1996-06-03 2000-01-25 The Johns Hopkins University School Of Medicine Method of detection of methylated nucleic acid using agents which modify unmethylated cytosine and distinguishing modified methylated and non-methylated nucleic acids
US6117635A (en) * 1996-07-16 2000-09-12 Intergen Company Nucleic acid amplification oligonucleotides with molecular energy transfer labels and methods based thereon
GB9812768D0 (en) * 1998-06-13 1998-08-12 Zeneca Ltd Methods
AUPP844899A0 (en) * 1999-02-01 1999-02-25 University Of Western Australia, The A method for detecting methylated nucleic acids
US6331393B1 (en) * 1999-05-14 2001-12-18 University Of Southern California Process for high-throughput DNA methylation analysis
US6277607B1 (en) 1999-05-24 2001-08-21 Sanjay Tyagi High specificity primers, amplification methods and kits
DE19951189C2 (en) * 1999-10-15 2003-11-06 Epigenomics Ag Method to distinguish 5-position methylation changes from cytosine bases and cytosine to thymine mutations and to detect single nucleotide polymorphisms (SNPs) or point mutations in genomic DNA
EP1278893A2 (en) 2000-04-06 2003-01-29 Epigenomics AG Diagnosis of diseases associated with dna replication
DE10029915B4 (en) 2000-06-19 2005-06-16 Epigenomics Ag Method for the detection of cytosine methylations
DE10112515B4 (en) 2001-03-09 2004-02-12 Epigenomics Ag Method for the detection of cytosine methylation patterns with high sensitivity
EP1409730A2 (en) * 2001-06-14 2004-04-21 University of Wales College of Medicine Virus detection method, primers therefor and screening kit
DE10132212B4 (en) * 2001-06-27 2005-11-24 Epigenomics Ag Method for the detection of cytosine methylation by comparative analysis of the single strands of amplicons
DE10154317B4 (en) 2001-10-26 2005-06-09 Epigenomics Ag Method for the detection of cytosine methylations in immobilized DNA samples
WO2005012569A1 (en) * 2003-08-01 2005-02-10 The University Of Western Australia Methods and kits for predicting the likelihood of successful treatment of cancer
US20060183128A1 (en) * 2003-08-12 2006-08-17 Epigenomics Ag Methods and compositions for differentiating tissues for cell types using epigenetic markers
WO2005098035A2 (en) 2004-04-06 2005-10-20 Epigenomics Ag Method for the quantification of methylated dna
EP3061833B1 (en) 2004-09-30 2018-03-14 Epigenomics AG Method for providing dna fragments derived from an archived sample
ES2546848T3 (en) * 2006-03-10 2015-09-29 Epigenomics Ag A method to identify a biological sample for methylation analysis

Also Published As

Publication number Publication date
JP2007502126A (en) 2007-02-08
JP4743787B2 (en) 2011-08-10
EP1835037A1 (en) 2007-09-19
US9863001B2 (en) 2018-01-09
US20060194208A1 (en) 2006-08-31
EP1654388B1 (en) 2007-06-27
EP1654388A2 (en) 2006-05-10
ES2289594T3 (en) 2008-02-01
DE10338308A1 (en) 2005-03-24
DE10338308B4 (en) 2006-10-19
WO2005024056A3 (en) 2005-06-02
CA2540378A1 (en) 2005-03-17
DK1654388T3 (en) 2007-10-29
WO2005024056A2 (en) 2005-03-17
ATE365813T1 (en) 2007-07-15

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